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目的:通过下调Lewis肺癌细胞中Vav3的表达,观察其对肺癌细胞体外迁移和侵袭能力的影响,探讨Vav3在肺癌转移过程中发挥的作用。方法:体外化学合成Vav3特异性siRNA(Vav3-163),并转染至Lewis肺癌细胞。蛋白质印迹法检测转染前后细胞中Vav3表达水平。Transwell小室实验观察下调Vav3对Lewis肺癌细胞体外迁移和侵袭能力的影响。结果:蛋白质印迹检测结果显示,转染Vav3-163序列后,Lewis肺癌细胞中Vav3的表达明显下调。实验组Vav3表达(0.112±0.005)低于LLC组(0.408±0.004)和对照组(0.388±0.005),F=22.052,P=0.002。下调Vav3在Lewis肺癌细胞中的表达后,迁移实验中干扰组穿过膜细胞数(31.0±10.5)明显少于Lewis肺癌细胞组(56.0±5.4)和对照组(49.0±9.1),F=11.213,P=0.002;体外侵袭实验中干扰组穿过人工基底膜细胞数(21.0±6.6)明显少于Lewis肺癌细胞组(59.0±9.8)和对照组(43.0±18.0),F=11.692,P=0.002。结论:下调Lewis肺癌细胞中Vav3的表达,具有抑制肺癌细胞体外迁移和侵袭能力的作用。
OBJECTIVE: To investigate the effect of Vav3 on lung cancer metastasis by down-regulating the expression of Vav3 in Lewis lung carcinoma cells and observing its effect on the migration and invasion ability of lung cancer cells in vitro. Methods: Vav3 specific siRNA (Vav3-163) was synthesized in vitro and transfected into Lewis lung cancer cells. Western blotting was used to detect the expression of Vav3 in cells before and after transfection. Effect of down-regulation of Vav3 on Lewis lung carcinoma cell migration and invasion in vitro by Transwell chamber assay. Results: Western blotting showed that Vav3 expression in Lewis lung cancer cells was significantly down-regulated after Vav3-163 transfection. The expression of Vav3 in experimental group (0.112 ± 0.005) was lower than that in LLC group (0.408 ± 0.004) and control group (0.388 ± 0.005), F = 22.052, P = 0.002. After the expression of Vav3 was down-regulated in Lewis lung cancer cells, the number of transmembrane cells (31.0 ± 10.5) in Lewis lung cancer cells was significantly lower than that in Lewis lung cancer cells (56.0 ± 5.4) and control group (49.0 ± 9.1) , P = 0.002. In vitro invasion assay, the number of cells passing through the artificial basement membrane in the interference group (21.0 ± 6.6) was significantly lower than that in the Lewis lung cancer group (59.0 ± 9.8) and the control group (43.0 ± 18.0), F = 11.692, P = 0.002. Conclusion: The down-regulation of Vav3 expression in Lewis lung carcinoma cells can inhibit the migration and invasion of lung cancer cells in vitro.